LOCUS       sequence-194327-        3044 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Rp51a with a tGGTAtGTta->AGGTAAGTAT 5'SS mutant with potential to 
            form 10bps with the U1 snRNA. Also contains TACTAAC->GTTAGTG BP 
            mutataion and a TACAAAC->GTTTGTG pseudo BP mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3044)
  AUTHORS   Larson JD, Hoskins AA
  TITLE     Dynamics and consequences of spliceosome E complex formation.
  JOURNAL   Elife. 2017 Aug 22;6. pii: 27592. doi: 10.7554/eLife.27592.
  PUBMED    28829039
REFERENCE   2  (bases 1 to 3044)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2017-08-22"; volume: "6. pii: 27592. doi"; pages: " 
            10.7554/eLife.27592"
COMMENT     Sequence Label: Rp51a_10bp 5'SS_No BP or pBP_pUC18
FEATURES             Location/Qualifiers
     source          1..3044
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(28..50)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     150..169
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     363..385
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     377..394
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     378..394
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     419..438
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        419..437
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(822..838)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(822..838)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(835..857)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    846..862
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(870..876,877..894,895..900))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    915..936
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1053..1070)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1224..1812)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1304..1323)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1983..2774,2775..2843))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2606..2625
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2844..2948)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3016..3034
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
       61 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
      121 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
      181 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
      241 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
      301 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
      361 ttcccagtca cgacgttgta aaacgacggc cagtgccaag cttgattacg ccaagctcta
      421 atacgactca ctatagggga aagctagctt gggatctcga gactagcaat aacaaaaagg
      481 taagtatata tggactaaag gaggctttta aggacacgta atattgagtc gaccgtgttt
      541 ttgatatcag tagttagtga agttgaattg cattgtttgt gtttttattt tgtattgctt
      601 ttcgtcattt taatagggta gagttagaac caagaccgtc aagcgtgctt ctaaggcttt
      661 gattgaacgt tactatccaa agttgacttt ggatttccaa accaacaaga gactttgtga
      721 tgaaatcgcc actatccaat ccaagagatt gagaaacaag attgctggtt acaccaccca
      781 tttgatgaag agaatccaaa agggtcggaa ttcgtaatca tgtcatagct gtttcctgtg
      841 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
      901 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
      961 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     1021 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     1081 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1141 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1201 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1261 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1321 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1381 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1441 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1501 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1561 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1621 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1681 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1741 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1801 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1861 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     1921 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     1981 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     2041 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     2101 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     2161 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     2221 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     2281 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     2341 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     2401 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     2461 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     2521 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     2581 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     2641 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     2701 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     2761 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     2821 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     2881 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     2941 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     3001 acattaacct ataaaaatag gcgtatcacg aggccctttc gtct
//
